A RAPID QUANTITATIVE DYE-BINDING METHOD OF SCREENING GLYCOSAMINOGLYCANS PRESENCE IN MEDICINAL PLANTS

Authors

  • Che Nur Mazadillina Zahari Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.
  • Marina Mohd Sham Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.
  • Sakina Shahabudin Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.
  • Mohd-hairul Ab Rahim Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.
  • Nina Suhaity Azmi Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.

DOI:

https://doi.org/10.22159/ajpcr.2019.v12i1.30283

Keywords:

Blyscan assay, Glycosaminoglycan, Medicinal plants, Orthosiphon stamineus, Plant-based source

Abstract

Objectives: The aims of this paper are to extract glycosaminoglycan (GAG) from four local medicinal plants and to characterize the crude extract with highest total sulfated GAG to reduce the dependency of using animals as major sources.

Methods: Crude GAG was extracted from four plants (Gaultheria procumbens, Strobilanthes crispus, Orthosiphon stamineus, and Ficus deltoidea) using hot water extraction with some modifications. Ultraviolet (UV) spectrophotometry was conducted for purity test. Total sulfated GAG was determined using Blyscan assay kit. By comparing results between the extract yields and total sulfated GAG, the plant consisting of high total sulfated GAG was chosen for further characterization. The selected plant sample was examined by microscopy and further analyzed by nuclear magnetic resonance (NMR) and Fourier-transform infrared (FTIR) spectroscopy.

Results: All four plants showed absorbance peaks between 214 and 232 nm in UV scan that represented negatively charged sugar. O. stamineus was found to contain the highest amount of sulfated GAG, 62.63±0.01 μg/mg by Blyscan assay. Microscopical examination confirmed the identity of O. stamineus sample by comparing to the reference. Both NMR and FTIR analysis of O. stamineus crude yield showed the presence of hydroxyl, sulfates, carboxylate, and amine groups, suggesting close resemblances to GAG structure.

Conclusion: The results suggested that all four plants contained GAG compound. O. stamineus was found to exhibit the most abundant total sulfated GAG and has the potential to become a new plant-based source for GAG.

Downloads

Download data is not yet available.

Author Biographies

Che Nur Mazadillina Zahari, Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.

Master student, Industrial Biotechnology Program, Faculty of Industrial Sciences and Technology UMP

Marina Mohd Sham, Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.

Industrial Biotechnology Program, Faculty of Industrial Sciences and Technology UMP

Sakina Shahabudin, Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.

PhD student, Industrial Biotechnology Program, Faculty of Industrial Sciences and Technology UMP

Mohd-hairul Ab Rahim, Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.

Lecturer, Industrial Biotechnology Program, Faculty of Industrial Sciences and Technology UMP

Nina Suhaity Azmi, Department of Industrial Biotechnology, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, 26300 Kuantan, Pahang, Malaysia.

Senior Lecturer, Industrial Biotechnology Program, Faculty of Industrial Sciences and Technology UMP

References

Sham MM, Azmi NS, Ab Rahim MH. Review: Glycosaminoglycans (GAGs) versus cancer. J Environ Bioremediat Toxicol 2014;2:58-61.

Zahari CN, Azmi NS. Halal source of medication: Glycosaminoglycan derived medicinal plant. Int J Appl Chem 2016;12:803-10.

Azmi NS, Fernig DG. Heparan Sulfate Surfaces to Probe the Functions of the Master Regulator of the Extracellular Space. Handbook of Biofunctional Surfaces. Singapore: Pan Stanford Publishing; 2013. p. 591-616.

Idrus NH, Azmi NS, Palliah JV. Waste to wealth: Alternative source of glycosaminoglycans (GAGs) from sea food waste. IIOAB J 2016;7 Suppl 1:543-7.

Sham MM, Azmi NS. Blue-spotted stingray: A promising source of beneficial glycosaminoglycans (GAGs). Int J Appl Chem 2016;12:795-802.

Lokwani R, Azmi NS, Yusoff MM, Ichwan SJ. Beyond anticoagulant: Heparin as a potential anti-cancer agent. J Biochem Microbiol Biotechnol 2014;2:76-82.

Toppo FA, Pawar RS. Novel drug delivery strategies and approaches for wound healing managements. J Crit Rev 2015;2:12-20.

Cesaretti, M, Luppi E, Maccari F, Volpi N. Isolation and characterization of a heparin with high anticoagulant activity from the clam Tapes phylippinarum: Evidence for the presence of a high content of antithrombin III binding site. Glycobiology 2004;14:1275-84.

Doughari JH. Phytochemicals: Extraction Methods, Basic Structures and Mode of Action as Potential Chemotherapeutic Agents. Phytochemicals-A Global Perspective of Their Role in Nutrition and Health. Croatia: INTECH Open Access Publisher; 2012. p. 1-32.

Azmir J, Zaidul IS, Rahman MM, Sharif KM, Mohamed A, Sahena F, et al. Techniques for extraction of bioactive compounds from plant materials: A review. J Food Eng 2013;117:426-36.

Azwanida NN. A review on the extraction methods use in medicinal plants, principle, strength and limitation. Med Aromat Plants 2015;4:196.

Choocheep K, Saenkham C, Borwornchaiyarit S, Chaiisaraseree K, Chimchai J, Moonpa T. Investigation of sulfated glycosaminoglycans and their agarose gel electrophoresis patterns from plants extracts. J Assoc Med Sci 2018;51:11-8.

Manjusha KP, Saleena M. Isolation and Characterization of Glycosaminoglycans and A Study of Its Bioactive Potential in Two Commercially Important Species of Cephalopods, Loligo duvauceli and Sepia pharaonis. A Thesis. Kochi: Cochin University of Science and Technology; 2011.

Movahedi A, Basir R, Rahmat A, Charaffedine M, Othman F. Orthosiphon stamineus: An Asian tea with substantial anticancer properties. J Nutr Sci Diet 2015;1:44-52.

Lima MA, Rudd TR, de Farias EH, Ebner LF, Gesteira TF, de Souza LM, et al. A new approach for heparin standardization: Combination of scanning UV spectroscopy, nuclear magnetic resonance and principal component analysis. PLoS One 2011;6:e15970.

Keng CK, Siong LP. Morphological similarities and differences between the two varieties of cat’s whiskers (Orthosiphon stamineus Benth.) grown in Malaysia. Int J Bot 2006;2:1-6.

Longas MO, Cheairs K, Puchalski MM, Park JI. Reliability of Fourier transform infrared spectroscopy in the characterization of human skin. Adv Biol Chem 2011;1:24-8.

Vidhyanandhini R, Saravanan R, Vairamani S, Shanmugam A. The anticoagulant activity and structural characterization of fractionated and purified glycosaminoglycans from venerid clam Meretrix casta (Chemnitz). J Liq Chromatogr Related Technol 2014;37:917-29.

Cael JJ, Isaac DH, Blackwell J, Koenig JL. Polarized infrared spectra of crystalline glycosaminoglycans. Carbohydr Res 1976;50:169-79.

DeAngelis PL, Gunay NS, Toida T, Mao WJ, Linhardt RJ. Identification of the capsular polysaccharides of Type D and F Pasteurella multocida as unmodified heparin and chondroitin, respectively. Carbohydr Res 2002;337:1547-52.

Guan D, Zhang Z, Yang Y, Xing G, Liu J. Immunomodulatory activity of polysaccharide from the roots of Actinidia Kolomikta on macrophages. Int J Biol 2011;3:3-10.

Widyaningsih TD, Rukmi WD, Sofia E, Wijayanti SD, Wijayanti N, Ersalia R, et al. Extraction of glycosaminoglycans containing glucosamine and chondroitin sulfate from chicken claw cartilage. Res J Life Sci 2017;3:181-9.

Singh N. A Comparison of Both Water and Ethanol Extracts Prepared from Echinacea purpurea and Echinacea angustifolia on the Response to Influenza A/PR/8/34 Infection in Mice. A Thesis. Ames, IA: Iowa State University; 2010.

Kee NL, Mnonopi N, Davids H, Naudé RJ, Frost CL. Antithrombotic/anticoagulant and anticancer activities of selected medicinal plants from South Africa. Afr J Biotechnol 2008;7:217-23.

Vergara-Salinas JR, Bulnes P, Zúñiga MC, Pérez-Jiménez J, Torres JL, Mateos-Martín ML, et al. Effect of pressurized hot water extraction on antioxidants from grape pomace before and after enological fermentation. J Agric Food Chem 2013;61:6929-36.

Barbosa I, Garcia S, Barbier-Chassefière V, Caruelle JP, Martelly I, Papy-García D, et al. Improved and simple micro assay for sulfated glycosaminoglycans quantification in biological extracts and its use in skin and muscle tissue studies. Glycobiology 2003;13:647-53.

de Jong JG, Wevers RA, Laarakkers C, Poorthuis BJ. Dimethylmethylene blue-based spectrophotometry of glycosaminoglycans in untreated urine: A rapid screening procedure for mucopolysaccharidoses. Clin Chem 1989;35:1472-7.

Linharattanaruksa P, Srisuwatanasagul S, Ponglowhapan S, Khalid M, Chatdarong K. Collagen and glycosaminoglycan profiles in the canine cervix during different stages of the estrous cycle and in open- and closed-cervix pyometra. J Vet Med Sci 2014;76:197-203.

Li WJ, Tuli R, Okafor C, Derfoul A, Danielson KG, Hall DJ, et al. A three-dimensional nanofibrous scaffold for cartilage tissue engineering using human mesenchymal stem cells. Biomaterials 2005;26:599-609.

Brown MP, Trumble TN, Sandy JD, Merritt KA. A simplified method of determining synovial fluid chondroitin sulfate chain length. Osteoarthritis Cartilage 2007;15:1443-5.

Davies Cde L, Berk DA, Pluen A, Jain RK. Comparison of igG diffusion and extracellular matrix composition in rhabdomyosarcomas grown in mice versus in vitro as spheroids reveals the role of host stromal cells. Br J Cancer 2002;86:1639-44.

Bahrom NA, Sirajudeen K, Yip GW, Latiff AA, Ghazali FC. Sulfated glycosaminoglycans from crown-of-thorns Acanthaster planci-extraction and quantification analysis. Food Sci Nutr 2013;1:83-9.

Ganesh EA, Das S, Arun G, Balamurugan S, Raj RR. Heparin like compound from green alga Chaetomorpha antennina-as potential anticoagulant agent. Asian J Med Sci 2009;1:114-6.

Tan HL, Chan KG, Pusparajah P, Lee LH, Goh BH. Gynura procumbens: An overview of the biological activities. Front Pharmacol 2016;7:52.

Adam Z, Hamid M, Ismail A, Khamis, S. Effect of Ficus deltoidea aqueous extract on blood glucose level in normal and mild diabetic rats. Malaysian J Health Sci 2007;5:9-16.

Misbah H, Aziz AA, Aminudin N. Antidiabetic and antioxidant properties of Ficus deltoidea fruit extracts and fractions. BMC Complement Altern Med 2013;13:118.

Murni A, Hanif N, Kita M, Darusman LK. Methyl 10-epi-pheophorbide A from MCF-7 cells active layer of the Indonesian Ficus deltoidea Jack leaves. Int J Pharm Pharm Sci 2017;9:183-6.

Fadzelly AB, Asmah R, Fauziah O. Effects of Strobilanthes crispus tea aqueous extracts on glucose and lipid profile in normal and streptozotocin-induced hyperglycemic rats. Plant Foods Hum Nutr 2006;61:7-12.

Olah NK, Radu L, MogoÅŸan C, Hanganu D, Gocan S. Phytochemical and pharmacological studies on Orthosiphon stamineus Benth. (Lamiaceae) hydroalcoholic extracts. J Pharm Biomed Anal 2003;33:117-23.

Pratiwi SU, Lagendijk EL, Hertiani T, Weert SD, Cornellius AM, Hondel JJ. Antimicrobial effects of Indonesian medicinal plants extracts on planktonic and biofilm growth of Pseudomonas aeruginosa and Staphylococcus aureus. Int J Pharm Pharm Sci 2015;7:183-91.

Akowuah G, Ismail Z, Norhayati I, Sadikun A. The effects of different extraction solvents of varying polarities on polyphenols of Orthosiphon stamineus and evaluation of the free radical-scavenging activity. Food Chem 2005;93:311-7.

Pan Y, Abd-Rashid BA, Ismail Z, Ismail R, Mak JW, Pook PC, et al. In vitro modulatory effects of Andrographis paniculata, Centella asiatica and Orthosiphon stamineus on cytochrome P450 2C19 (CYP2C19). J Ethnopharmacol 2011;133:881-7.

Yam MF, Ang LF, Basir R, Salman IM, Ameer OZ, Asmawi MZ, et al. Evaluation of the anti-pyretic potential of Orthosiphon stamineus benth standardized extract. Inflammopharmacology 2009;17:50-4.

Yam MF, Lim V, Salman IM, Ameer OZ, Ang LF, Rosidah N, et al. HPLC and anti-inflammatory studies of the flavonoid rich chloroform extract fraction of Orthosiphon stamineus leaves. Molecules 2010;15:4452-66.

Kato-Noguchi H, Hamada N, Morita M, Suenaga K. A novel allelopathic substance, 13-epi-orthosiphol N, in Orthosiphon stamineus. J Plant Physiol 2013;170:1-5.

Takeda Y, Matsumoto T, Terao H, Shingu T, Futatsuishi Y, Nohara, T, et al. Orthosiphol D and E, minor diterpenes from Orthosiphon stamineus. Phytochemistry 1993;33:411-5.

Naumann D, Helm D, Labischinski H. Microbiological characterizations by FT-IR spectroscopy. Nature 1991;351:81-2.

Published

07-01-2019

How to Cite

Zahari, C. N. M., M. M. Sham, S. Shahabudin, M.- hairul Ab Rahim, and N. S. Azmi. “A RAPID QUANTITATIVE DYE-BINDING METHOD OF SCREENING GLYCOSAMINOGLYCANS PRESENCE IN MEDICINAL PLANTS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 12, no. 1, Jan. 2019, pp. 396-02, doi:10.22159/ajpcr.2019.v12i1.30283.

Issue

Section

Original Article(s)